RU2680677C2 - Plasmapheresis device - Google Patents

Abstract

FIELD: medicine.SUBSTANCE: invention relates to medicine, specifically to efferent therapy, and can be used during plasmapheresis. For this purpose, a device for plasmapheresis is proposed, which comprises a first conduit through which blood drawn from a mammalian patient travels under the influence of a pump, a separator for separating cellular components of said blood from plasma, a column device through which said plasma flows. Column device selectively removes galectin-3 (gal-3) and other substances associated with galectin-3, including TNF receptor blockers from said plasma to provide treated plasma. Device also comprises a second conduit, which receives said treated plasma after passing through said column device and in which said plasma to be treated is combined with blood cells separated in said separator before returning to said patient. Said first and second conduits provide for a substantially continuous flow of said blood and plasma from said patient and return to said mammal. Device also comprises at least one additional column device for removing a plasma component other than gal-3.EFFECT: disclosed device provides not only the removal of galectin-3 or other galectins from blood, but also the removal of other, including potentially dangerous, factors and also provides for the introduction into the blood of additional therapeutic factors when returning the blood to the mammalian patient.14 cl, 3 dwg

Description

BACKGROUND

FIELD OF THE INVENTION

[0001] The present invention is directed to devices for performing apheresis of mammalian blood, including the selective removal of a certain percentage of circulating galectin-3 (gal-3) from the blood. The device has the ability not only to remove galectin-3 (or other galectins) from the blood, but other potentially harmful factors, and to introduce additional factors into the blood that are therapeutic or beneficial when blood is returned to the patient to the mammal.

BACKGROUND OF THE INVENTION

[0002] This invention relates to the subject matter disclosed and claimed in the United States Patent Application Serial Number 13/863989, filed September 28, 2012, and 14/141509, filed December 17, 2013. The disclosure of both of these cases is incorporated herein by reference. With legal permission, the Applicant claims the priority of the filing date of these two applications.

[0003] Galectin-3 was the focus of a number of studies aimed at explaining the mechanisms that contribute to the onset of inflammation, fibrosis, and various forms of cancer in mammals. Under “mammals” in this application, the applicant intends to describe, of course, humans, but also mammals of sufficient value, worthy of investment in their health, including domestic mammals such as dogs and cats, and mammals of industrial importance, such as cows, horses, goats and pigs. Higher-order mammals, such as monkeys, which can serve as research models, as well as pets, are also included in this term. The invention, of course, is focused on people.

[0004] Galectins are a family of lectins (sugar binding proteins) that are characterized by the presence of at least one carbohydrate-recognition domain (CRD) with affinity for beta-galactosides. These proteins were recognized by the family only recently, but were found in the entire animal kingdom, and were found in mammals, birds, amphibians, fish, sponges, nematodes, and even fungi.

[0005] Galectins mediate and modulate a wide range of intracellular and extracellular functions, and thus are both expressed within the cell and often target a specific cytosol region and are secreted from the cell for distribution outside the cells as a component of human plasma. Among the many functions that mediate extracellular galectins, there is inflammation, fibrosis, cell adhesion, cell proliferation, metastasis, angiogenesis (cancer), and immunosuppression.

[0006] Galectins are a family of fifteen (15) carbohydrate-binding proteins (lectins) that are highly conserved for all animal species. Most of the galectins are widespread, although galectin-5, -10 and -12 exhibits tissue-specific distribution. While galectins are variablely expressed by all immune cells, they have an increased level of expression in activated B and T cells, inflammatory macrophages, natural killer (NK) cells, and FoxP3 regulatory T cells. Galectins contain a number of structural configurations, but a relatively conservative carbohydrate-recognizing domain (CRD). Most galectins exhibit a single CRD, and are biologically active as monomers (galectin-5, -7 and -10), or require homodimerization for functional activity (galectin-1, -2, -11, -13, -14 and -15 ) Alternatively, galectins (galectin-4, -8, -9 and -12) of the tandem repeat type contain two CRDs separated by a short linker peptide, while galectin-3 (chimeric type) has a single CRD fused to a non-lectin domain that may be in the form of a complex with other galectin-3 monomers to form an oligomeric pentamer. It is noteworthy that some galectins, such as galectin-10, bind to mannose-containing glycans. Among the galectin family, -1, -3, and -9 are the most important as potential therapeutic targets, and -2, -4, -5, -6, -7, -8, -10, -11, -12, - 13, - 14 and -15, apparently, are also involved in a number of biological metabolic pathways associated with morbidity and mortality.

[0007] Thus, galectin-7 is involved in the development of certain forms of cancer, St. Pierre et al, Front. Biosci., 1:17, 438-50 (2012), and in a number of specific forms of cancer, including gal-2, -4, and -8 in connection with colon and breast cancer, Barrow et al, Clin. Cancer Res, 15; 17 (22) 7035-46 (2011). Squamous cell carcinoma of the tongue, Alves et al, Pathol. Res. Pract. 15; 207 (4) 236-40 (2011) has been shown to be associated with elevated gal-1, -3, and -7 levels, while squamous cervical carcinoma of the cervix has been shown to be associated with gal-7 levels, Zhu et al, Int. J. Cancer, (Aug., 2012). It has been demonstrated that a number of galectins, including gal-15, gal-13 and gal-10, are associated with implantation and pregnancy problems. See, e.g., Than et al, Eur. J. Biochem. 271 (6) 1065-78 (2004), Lewis et al, Biol. Reprod. 77 (6); 1027-36 (2007). A number of galectins, including gal-2, 3, 8, and others, have been identified as correlating with various autoimmune disorders, such as lupus. Salwati et al, /. Infect. Dis. one; 202 (1) 117-24 (2010), Pal et al, Biochim. Biophys. Acta., 1820 (10) 1512-18 (2012) and Janko et al, Lupus 21 (7): 781-3 (2012). Elevated levels of a number of galectins, including gal-3, are associated with inflammation and fibrosis, which occurs during wound healing and the like. Gal et al, Acta. Histochem. Cytochem. 26:44 (5); 191-9 (2011). It is clear that the mediation of inflammatory and fibrous metabolic pathways makes galectins the most important elements of a wide range of diseases, injuries and phenomena associated with trauma. In many cases, the presence of undesirable concentrations of galectins can aggravate a disease state or trauma situation, or prevent attempts to treat diseases such as cancer or congenital heart disease. Among the galectin family recognized as active in humans, galectin-1, galectin-3 and galectin-9 are of particular interest. As indicated above, these proteins are commonly referred to and are referred to herein as gal-1, gal-3 and gal-9. A wide range of pathological conditions in people, ranging from conception problems to asthma, chronic heart failure, cancer, viral infection, stroke and not only, are mediated or exacerbated by higher than normal concentrations of galectins. Thus, among other galectins, gal-3 is especially prominent in fibrosis, inflammation, and cell proliferation, whereas gal-1 also plays a role in the immunosuppression necessary for a successful pregnancy. It is believed that Gal-1 is also involved in the differentiation of nerve cells. It was shown that Gal-9 is involved in the regulation of lesions resulting from immuno-inflammatory diseases, is generally involved in inflammation - gal-9 undoubtedly plays a role in replenishing eosinophils in areas of inflammation. Apparently, it also mediates apoptosis in certain activated cells. While the discussion in this application is applicable to circulating active gal-1, gal-3 and gal-9 and to galectins in general, when elevated levels of circulating galectin are associated with a disease or damage conditions, more was found out about the role of gal-3 in progression diseases and injuries than any of the other galectins, and therefore is given as an example in this document. More specifically, the invention is focused on the removal of active gal-3 from the plasma of a mammal, in particular humans. It was demonstrated that Gal-3 is involved in a large number of biological processes, many of which relate to various types of disease states. The binding and blocking activity of gal-3 in the circulating blood, or the removal of large quantities of gal-3 from the blood circulation can thus improve existing treatments, suppress and / or reduce inflammation and fibrosis, which are the result of others, and provide the possibility of intervention in various painful conditions, not otherwise amenable to easy treatment. The invention is equally applicable to lowering the circulating levels of other active galectins to resolve pathological conditions mediated by these galectins. What is referred to as “active” galectins is biologically active molecules. As noted, for example, gal-3 can be active, that is, mediate mammalian responses to various injuries and pathological conditions, in the form of a monomer and in the form of an oligomer. In any mammal, at any given time, significant amounts of gal-3 and other galectins are inactive — that is, they are either bound to tissue or bound to a ligand so as to inhibit molecular interaction. Despite the fact that such galectin molecules can become active and can or become a target for removal by the invention disclosed herein, by monitoring patient pathological conditions and managing responses, the focus of the invention is a device suitable for removing active galectins from the bloodstream. The present invention provides the use of plasmapheresis, sometimes referred to as apheresis, and / or replacement of therapeutic plasma, for controlling blood levels of gal-3, and more particularly biologically active galectin. First, whole blood is divided into cellular and plasma components. The plasma is then passed along the fluid path and is either mixed with a gal-3 binding agent that can be isolated from the plasma, either returned to the blood with blocked inactivated gal-3, or passed through a solid substrate that binds gal-3, this in subsequent plasma is returned to the body with a reduced level of gal-3. Thus, the present invention can be used to remove bound gal-3 as part of a strategy to reduce the total gal-3 content. However, the focus in this application is to remove, as a therapeutic measure, active or unbound gal-3.

State of the art

[0008] This application relates to US Patent Application Serial No. 13/153648, filed June 6, 2011. In turn, that application claims the benefit of priority of US Patent 8,426,567, issued April 23, 2013. The contents of both of these patent documents are expressly incorporated herein by reference. US Patent Application Serial No. 13/153648 (US Patent Publication US-2011-0294755 Al) discloses a method for treating pathological conditions of cell proliferation, inflammation and enhanced fibrosis, which comprises administering an agent that can bind circulating gal-3, such as modified citrus pectin, (MCP), citrus pectin that has a reduced molecular weight of twenty thousand (20,000) daltons or less, preferably ten thousand (10,000) daltons or so. MCP is commercially available from EcoNugenics Santa Rosa, California and is discussed in US Pat. Nos. 6,274,566 and 6,462,029.

BACKGROUND OF THE INVENTION Biolo2V

[0009] Gal-3 is approximately 30 kDa and, like all galectins, contains a carbohydrate recognition and binding domain (CRD) of approximately one hundred thirty (130) amino acids, which is capable of specifically binding β-galactosides. Gal-3 is encoded by a single gene, LGALS3, located on chromosome 14, locus q21-q22. It has been demonstrated that this protein is involved in a large number of biological processes. The list set forth in this document is only illustrative as continuously identifying new situations and roles for gal-3. Among the biological processes at the cellular level, which have been shown to be gal-3 mediated at least partially, cell adhesion, cell migration, cell invasion, cell activation and chemo-attraction, cell growth and differentiation, angiogenesis and apoptosis are present.

[0010] It has been demonstrated that the wide biological functionality of this gal-3 is involved in a large number of disease states or medical complications. Studies have also shown that gal-3 expression is involved in a number of processes related to heart failure, including myofibroblast proliferation, fibrogenesis, tissue healing, inflammation and remodeling of the ventricles and tissues. It was found that elevated blood levels of gal-3 are significantly associated with increased morbidity and mortality. It was also found that they are significantly associated with a higher risk of death in populations with both acute decompensated heart failure and chronic heart failure.

[0011] Various studies have shown that elevated gal-3 levels exacerbate a wide range of disease conditions associated with cell proliferation. High gal-3 levels are associated with cancer growth and cancer progression to metastasis in a wide variety of cancers. A number of cancers are specifically associated with or associated with elevated gal-3 levels, including liver cancer, kidney cancer, breast cancer, prostate cancer, colon cancer, thyroid cancer, gall bladder cancer, nasopharyngeal cancer, lymphocytic leukemia, lung cancer, melanoma , multiple myeloma, glioblastoma multiforme, uterine cancer, ovarian cancer, cervical cancer, brain cancer and others. It has also been shown that elevated gal-3 levels inhibit or suppress conventional antitumor regimens, such as chemotherapeutic treatments such as cisplatin, doxorubicin and similar chemotherapeutic agents.

[0012] Inflammation is a frequently occurring condition of the body - a natural response of the body to a number of diseases and trauma. As with the other pathological conditions noted above, gal-3 levels above normal levels are involved in a wide variety of situations where fatal inflammation occurs. Once again, the list of pathological conditions and disease states is too extensive to exhaust every possibility, but inflammatory pathological conditions associated with elevated gal-3 levels include increased inflammation associated with non-degradable pathogens, autoimmune reactions, allergies, exposure to ionizing radiation, diabetes, heart dysfunction disease, arteriosclerosis, inflammation of the bronchi, ulcers of the gastrointestinal tract, inflammation of the colon, inflammation of the liver, associative associated with cirrhosis, inflammation associated with a parasitic infection, inflammation associated with a viral infection, inflammation associated with a fungal infection, inflammation associated with a bacterial infection, inflammation associated with infection by intracellular organisms and associated infections such as tuberculosis, sarocoidosis, feline disease scratches, mycoplasma, Lyme disease, bartonellosis, ehrlichiosis, rickettsial diseases, babesiosis and others. Inflammation associated with arthritis, multiple sclerosis, psoriasis, Alzheimer's disease, Parkinson's disease and amyotrophic lateral sclerosis (ALS) may also be considered. And again, although inflammation is a pathway often used by the body in response to any number of infections, it has been found that elevated gal-3 levels exacerbate and contribute to inflammation, causing lesions and injuries leading to widespread morbidity and mortality a number of situations that are otherwise manageable, including inflammation due to poisoning by heavy metals, pesticides, oxidizing agents, xenoestrogens and similar toxins, stroke and related ischemic injuries, acetaminophen-induced liver inflammation, a series of T-cell-mediated responses usually involved in autoimmune diseases, and so on. Gal-3 is also involved in kidney damage and kidney disease, hepatitis, pulmonary hypertension and fibrosis, diabetes and inflammatory conditions of the gastrointestinal tract, such as ulcerative colitis, Crohn’s disease, celiac disease and others.

[0013] As noted, elevated levels of circulating, active gal-3 are associated and undoubtedly contribute to a number of inflammatory pathological conditions, including conditions that contribute to diseases of the cardiovascular system, kidney, lung, brain, and liver. Gal-3 is also associated with the formation of fibrosis, in particular in response to organ damage. It has been found that higher levels of circulating gal-3 induce pathogenic fibrosis in cardiovascular disease, gastrointestinal tract disease, cardiovascular trauma, renal tissue trauma, brain injury, lung trauma, liver tissue trauma, tissue damage due to radiation therapy and diseases and pathological conditions of connective tissue and skin, such as systemic scleroderma.

[0014] Accordingly, the technical field is replete with observations that elevated levels of gal-3, as well as gal-1 and gal-9, can complicate or aggravate a wide range of diseases and conditions of damage. It would be valuable to find a way to regulate inflammation and the formation of fibrosis when inflammation and fibrosis are damaging, in particular in the environments described above, and in particular in cardiac care and disease and trauma to other organs and tissues. Similarly, it would be valuable to regulate gal-3 mediated cellular responses that accelerate cell proliferation and transformation, including tumor formation and growth, cancer cell transformation, and metastatic spread of cancer. Another objective in this area is to avoid the problems associated with the influence of elevated gal-3 levels in the treatment of cancer with conventional agents such as bleomycin, doxorubicin (adriamycin) and other anthracyclines, cyclophosphamide and cyclosporin, as well as targeted biological agents, such as VEGF and EGF inhibitors, examples are bevacizumab (Avastin) and erbitux (Cetuximab). Some of the side effects caused by these agents are gal-3 mediated and can be affected and improved by the invention, while their mechanism of action can be blocked by gal-3 (enhancing angiogenesis). Elevated gal-3 levels also appear to inhibit pharmaceuticals used in other uses, such as the antiarrhythmic drug amiodarone (cordarone) and statin drugs.

[0015] Plasmapheresis is a blood separation technique where blood is removed from the body through a needle or catheter to a separator that removes blood cells and returns them to the body, leaving the plasma. Cells return to the body when the plasma is reconnected with cellular components, or a replacement fluid has been added to the blood cells. This type of technique has historically been used in the treatment of autoimmune diseases, where the antibodies in question are removed by contacting the plasma with the ligands with which they bind. Then, if necessary, the plasma was supplemented with anticoagulants, drugs and related elements, recombined with blood cells, and returned to the body. In a prior art, methods involved plasma substitution or mostly replacement therapies, as illustrated in US Patent Publication US 2006/0129082, the technique being used to isolate and remove “toxic serum components” such as ammonia, uric acid and growth inhibitors cells. The same reference in [0009] - [0010] warns against the use of plasma substitution in general. Similar warnings were made by Kyles et al, Am. J. Crit. Care, 14, 109-112 (2005), who analyzed the use of plasmapheresis to help treat sepsis with immunoglobulins, noting that plasmapheresis has traditionally been used in treatment to remove pathogenic autoantibodies and endotoxins in autoimmune disorders and to remove harmful substances produced by infectious organisms that cause sepsis .

BACKGROUND OF THE INVENTION Plasmapheresis Devices

[0016] An early form of a plasmapheresis device is set forth in US Pat. No. 3,625,212, which describes measures to ensure return of treated plasma as well as separated blood cells to an appropriate donor. US Patent No. 4,531,932 teaches plasmapheresis by centrifugation, a method used to separate red blood cells on a fast and near-continuous basis. Each of US Pat. Nos. 6,245,038 and 6,627,151 describes a number of methods for separating the plasma contents and returning the treated plasma to the patient after the first removal of red blood cells, generally to reduce blood viscosity by removing high molecular weight protein. While the invention that is the subject of this application focuses on reducing levels of circulating galectins, such as levels of gal-3, and low molecular weight proteins, or directly affecting viscosity, the disclosure of these four (4) patents is incorporated herein by references for their disclosure of available plasmapheresis techniques and devices, which may be generally involved in this invention. The advantages of apheresis in general, including plasmapheresis, have demonstrated the efficacy of hemodialysis equipment using a highly permeable membrane similar to Asahi Medical's Plasmaflo AP-05H and a standard dialysis device in ultrafiltration mode. In application, this is similar to hemoperfusion.

SUMMARY OF THE INVENTION

[0017] The plasmapheresis device of the present invention improves the scope of the invention by selectively removing gal-3 from plasma, which is separated from whole mammalian blood if active gal-3 is to be reduced. As stated in US patent applications simultaneously filed, serial number 13/863989, filed September 28, 2012, and 14/141509, filed December 27, 2013, the removal of active gal-3 from the bloodstream of a mammal can affect the resolution of unwanted inflammation to reduce the risk. which is fibrosis, in particular cardiac fibrosis, to reduce the risk or spread of various types of cancer, to improve the types of treatment, etc. The decisive requirement in this plasmapheresis device, therefore, is the selective removal of gal-3. This depends on the passage of the separated plasma through the component that selectively binds gal-3. This can be a column, container or similar platform, which physically contains material that positively binds gal-3, for example, an antibody to it, so when plasma passes through it, a significant part of the gal-3 trapped in it is bound by a column. In other embodiments, selective removal of gal-3 is accomplished by combining the plasma stream in the machine tube with material that binds gal-3, and then is easily removed, either through an antibody specific to it, or through adjuvants such as attached magnetic particles that can be extracted by selective application of a magnetic charge.

[0018] However, the invention is not limited to the removal of gal-3. Ideal machines provide this selective removal ability with the ability to remove other components from the blood that can either mediate or exacerbate painful conditions. As just one example of a possible development of events, a machine can be equipped with multiple columns for removal or absorption, one removing gal-3, the second removing TNFa, and the third removing TGF, or with the same column having multiple filters in sequential order for removing these three. Additional examples of compounds to be removed are C-reactive protein (CRP), fibrinogen, NFkβ, various inflammatory cytokines, etc., which may be involved in a patient with chronic inflammation in a number of disease states, including cancer. Not only does the blood treatment in the proposed device remove enhancers that enhance the inflammatory response, but due to this, and possibly removing the receptors for these markers that block them, it is possible to improve the effectiveness of anticancer drugs or treatments. Thus, the device of the invention advantageously contains a plurality of columns or deletion elements for the selective removal of not only gal-3, but other agents with the most frequently detected increase in blood in individuals with a number of both chronic and acute pathological conditions.

[0019] Although, as also described in previous cases, effective treatment may not only depend on the selective removal of gal-3 from the blood and other blood agents, but it may depend on the addition of therapeutic or beneficial elements before the blood is returned to the mammal. Often, the addition of an agent to plasma is an effective and efficient means of prescribing a treatment, such as a drug, vitamin, or similar component. Thus, although this is not essential for the selective removal of gal-3, the device of the invention of the present invention includes multiple ports for adding a number of agents to the plasma and circulated whole blood. Examples include various homeopathic medicines, metabolic regulators, immune response modifiers, medicines and various cytotoxic compounds, antimicrobial agents, chelating agents, possibly targeted drugs, to assist in the treatment.

BRIEF DESCRIPTION OF THE DRAWINGS

[0020] The accompanying drawings, which are incorporated herein and form part of this description, illustrate exemplary embodiments of the present invention, and together with the general description above and the detailed description below, serve to explain the features of the invention.

FIG. 1 is a broad schematic representation of the device of the present invention, reflecting the separation of whole blood into blood and plasma, plasma treatment, the return of plasma to the blood component, and compensation of the treated whole blood to the patient.

FIG. 2 is a schematic representation of a portion of a device where plasma is separated from blood (which may occur through one of several existing separation techniques) processed in at least one column and returned through a tube to the blood and donor.

FIG. 3 is a diagram of the device of the invention, intentionally repeating the structure of FIG. 1, but reflecting the ability of this device to install multiple columns and ports for both removing and adding a number of components to the plasma and ultimately the patient.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

Description

[0024] The plasmapheresis device of the present invention is generally depicted in Figure 1. Blood is removed from a patient of a mammal 1002, usually through the insertion of a venous catheter, either peripheral into a limb or into a central vein. The central veins make possible higher blood flow velocities and are more convenient for repeated procedures, but are more often the site of complications, especially bacterial infections. A catheter is not part of the device of the invention per se, and is usually provided by an operator, such as a clinic or hospital. A catheter is connected to the device’s tube, 1000, and a forced flow, assisted by a pump 1006, distributes blood through the system.

[0025] As soon as the blood is essentially removed from the patient’s body, the plasmapheresis device starts to work in the usual way. Thus, as a first step in using the device, an anticoagulant such as heparin or a substance of artificial origin, such as acenocoumarol or phenindione, is provided through port 1004. Adding an anticoagulant effective to prevent thrombosis induced by current through the tube and returning to the patient is mandatory. For many individuals, a port will be provided between the addition of the anticoagulant and the blood / plasma separation device 1012 will be useful, as illustrated in Figure 1, but is not required.

[0026] In order to reduce viscosity and pressure and make the treatment practical and effective, cellular components such as red blood cells (red blood cells), white blood cells (WBC) and platelets are removed and separated from the plasma. This is carried out on a separation device 1012. The separation device may be any conventionally used. Centrifugal separation is an old process in the field of technology. It may include intermittent flow centrifugation, where a single venous catheter is used, and aliquots of blood, possibly up to about 500 ml, are removed and centrifuged simultaneously to remove blood cells from the plasma. Continuous flow centrifugation is also known, which reduces the amount of blood outside the body at any given time, but requires the use of two systems for intravenous infusion. This is the most commonly used method when blood is removed from one area and returned through another area, usually in two different limbs. The catheters used make return flow possible. A continuous flow is almost always used, with only 180 ml of blood outside the body (extracorporeally) at a certain point. This procedure usually draws an average of 60 ml / min of blood and can filter 1.5 times the plasma volume in about 2 hours. The flow rate and the volume of filtered plasma can be adjusted based on medical needs.

[0027] As an alternative to centrifugation, plasma membrane separation using now standard dialysis membranes may be softer, but may increase the processing time.

[0028] As a general observation throughout the use of the device of the invention, support and monitoring of the patient is required. Although the device can be largely automated, it is not a device generally suitable for home or uncontrolled use. To this end, an operator’s control station or interface that allows direct control, as well as automation of almost any function of the device, including agent levels, such as added anticoagulant, transport speed, column type, sampling port and the like. It is practical and possible to continuously monitor a wide range of mammalian parameters using the device, and ports for sampling are provided for this purpose. Part of the operating platform may include a monitoring device that monitors heart rate, oxygen saturation, the main rhythm of the ECG, and blood pressure.

[0029] In the separation unit 1012, a stream extracted from a mammalian patient 1002 is separated into red blood cells and other cellular components maintained in a return flow fluid. One or more ports are usually provided in this catheter prior to return. There may be a number of reasons for removing cells at a given point. It can be combined with a plasma separation device, but ports are provided to remove various cells in addition to separating blood cells, including cells such as stem cells, T cells, B cells, NK cells, etc. There is more opportunity to capture various cells from plasma and use or return, but this operation can be combined and integrated into block 1012 for plasma separation.

[0030] The plasma from the separator is directed by a pump 1010 through at least one column 1008. Column 1008 contains, as described above, one or more elements that selectively bind or remove gal-3 from the plasma. In the most traditional format, a “column” is a tube that is lined with a mechanically fixed binding agent, such as an antibody bound to the inner wall of the tube through an agent such as Sepharose, but a wide variety of antibody-binding proteins for in vitro plasma passage is well known. including Protein A, Protein G and Protein L. Other binders, such as negatively charged compounds, resins, glycoproteins, etc., can be used. As the plasma passes through the tube, it is necessary to “wash” the binding molecules that selectively remove a portion of gal-3 from the plasma. The columns may be of any known type and, as indicated above, may not actually be “traditional columns” (such as centrifuge columns), but may actually be open channels where gal-3 is selectively bound by modified particles, such as pectin particles, which are biocompatible with either a galectin-binding molecule, N-acetyl-lactosamine, or with a modified pectin, such as citrus pectin. These particles are typically modified to include an element that makes their fixation and / or removal effective, such as magnetically attracted particles, for binding with a suitable magnet. The nature of the column is not particularly limited except that it selectively binds gal-3. The size and number of columns will vary, and ideally the device of the invention can be adapted to a different number of columns based on the needs of the patient. A single column may be effective in removing, for example, 10% or more active gal-3 from a patient’s blood circulation. While many patients will have ten percent effective for the therapeutic effect, some patients may require greater removal, depending on the nature of the selection criteria, including, for example, selection to alleviate inflammation, as opposed to inhibiting the spread of metastatic cancer. To this end, the device of the invention can be provided with a plurality of columns 1008. Thus, the use of the device can be effective in removing fifteen percent, twenty percent or even more, up to possibly forty percent of circulating gal-3. The amount of gal-3 selectively removed by column (s) 1008 is actually limited by the hydrodynamics of the system, the affinity for binding materials, and the needs of the patient. regenerating columns can be placed in their place parallel to each other, so each can be regenerated or cleaned, and the machine can switch between them automatically at certain intervals of the plasma volume. Related features commonly used in plasmapheresis technology include discharge lines for whole blood, plasma, and columns, each having discharge filters and pressure control lines. Separate pumps are needed for whole blood, and then for the plasma path, and then also for certain types of columns, such as double regeneration adsorption columns, since they are washed and cleaned and alternated one after another, they need their own pump. Also widely used are air detectors, blood leakage detectors, conductivity detectors, blood heaters, waste bags / lines, etc. and fluid inlet lines for column regeneration. they are collectively referred to herein as column care items.

[0031] After plasma treatment, including selective removal of gal-3, the plasma stream is reconnected with the stream of cellular components (RBC, WBC, platelet), and the reconnected blood is returned to the patient. Often, in order to facilitate the return of the flow due to the addition of ports through the ports to the device’s tubes, some build-up level is used using synthetic plasma, albumin, auxiliary blood cells and sometimes saline. Observed speeds and pressures are common. Accordingly, the treatment may include several hours when the patient is connected to the machine. Effective treatment depends on a wide range of factors, including patient factors and the capabilities of the machine itself. Often processing occurs more than once a week, and in acute cases, more than once a day. the concentration of Gal-3, as well as the concentration of other agents targeted for the patient in question, are monitored both in the returned plasma / blood and in the extracted blood, in order to allow monitoring and equilibration. When the active levels of gal-3 decrease to a favorable level, the blood extraction is completed, and when the last plasma / blood is returned to the patient, the treatment is completed.

[0032] Figure 2 reflects the "plasma loop" of the device. As shown, the whole blood extracted from the patient, after adding the anticoagulant, is captured in the tube 2000. In Figure 2, the separation of blood / plasma occurs in the separation device 2012, which is not necessarily a membrane separation device, where the passage is allowed depending on the size. Plasma is diverted in the direction of the arrows, as shown, despite the fact that red blood cells and other cellular components continue to pass through the tube 2000. The principal feature of this loop is the “column” 2008. There is no need for the column to be a traditional column. Hollow fiberglass filter technology is widely available. A typical product is a product offered by Calux, China, but numerous suppliers are well known. New and distinctive in relation to this “column” is the selective removal of gal-3.

[0033] It is clear that a patient whose condition can be improved by plasmapheresis is most likely to receive additional therapeutic benefit from the removal of other abnormally elevated or potentially harmful substances or agents found in the plasma, and therefore more than one “column” can be used 2008. in preferred embodiments, the apparatus of the present invention can provide a plurality of “in-cassette” filters rotating in or out. Possible targets for co-removal along with selective removal of gal-3 may include heavy metals (e.g. mercury), antibodies such as dangerous autoantibodies, pesticides or toxins, LDL, Lp-PLA-2, LP (a), oxidized cholesterol oxidized LDL, cytokines (examples such as IL-6, IL-8), various types of immunoglobulins, complement, CRP, TNFa, receptors for TNFa, TGF, NH, growth factors such as VEGF, other inflammatory components such as CRP, fibrinogen, heavy metal binding proteins such as ceruloplasmin for copper, other galectins (in particular pebbles Ins-1 and 9) and cytokine receptors such as IL-2 receptors, TNF-a etc. Accordingly, the provided gal-3 selective removal is provided at the beginning, in the middle, or at the end of the plasma treatment loop, wherein other elements can be removed using columns specific for their removal.

[0034] As noted, in addition to the gal-3 removal value, the inventive device may advantageously include the addition of various agents before the blood is again administered to the patient. Supplements are described above, as well as other drugs. As one separate illustration, the disclosure included makes it clear that removing a certain amount of circulating gal-3 can increase the effectiveness of certain anti-tumor pharmaceuticals. To maximize the increase, the pharmaceutical preparation should preferably be added to the plasma or reunited blood stream before returning to the patient, thus adding at a time when the patient's gal-3 levels seem to be the lowest. An optimized device is illustrated in Figure 3, so that a person skilled in the art can compare the simplified device with the above description. Blood from patient 3002 flows again through a tube 3000 where an anticoagulant has been added to prevent ex vivo blood clot formation, as well as after reverse administration to the patient. An anticoagulant is added in 3004, after which blood flows through a pump 3006, past port 3016. Many ports 3016 are included in the tubes through which blood, red blood cells and plasma flow. Three ports are illustrated, but in fact, the device can contain as much as acceptable for the purpose, and will usually contain more. Six may be a more realistic value, but the ports can be used both to dispense fluid and to add the elements discussed above. In preferred embodiments, port control, including withdrawal and addition, is performed by an automated device management system, with the operator preselecting operating options, optionally with a manual control option. Ports are predominantly provided in the catheter through which the blood of the patient is discharged, the plasma catheter, the red blood cell catheter after separation, and in the combination catheter before being reintroduced to the patient, but the placement and selection of ports will vary from machine to machine.

[0035] In addition to removing and adding the various elements discussed above, the described device allows the use of separators, such as a separator 3012 to "take" a number of cells for treatment or use, such as B cells, T cells, platelets, stem cells etc. Many of these cells and fragments are effectively preserved for later treatment of the patient. Advantageously, the cells can be removed and modified and returned by the same device, for example, T-cell or B-cell lymphocytes processed to be able to recognize cancer cells of a particular type in a particular patient and attack them. When reintroduced into this device, while gal-3 and other inflammatory mediators decrease, the antitumor effect may be enhanced while avoiding a dangerous and often life-threatening inflammatory response. Various types of reinforcing therapies, such as CTC (tumor cell circulating in the blood) -therapy, receive the greatest advantage through this type of device. Among the several advantages of using this machine are simplified sampling of the patient, a reduced chance of infection and improved monitoring and control. In CTC, tumor cells are removed from the separated plasma in a separator. These cells, which are essentially metastatic tumor emboli, are modified to carry antibodies and / or viruses and returned to the body. In fact, they become selectively destroying cells that the body recognizes and does not attack. They can also be used to create antibodies that are then introduced back into the body and attack any cancer present in the patient, or the lymphocytes taken from the patient with cancer are sensitized to certain surface membrane proteins specific to the patient’s cancer cells and then returned. Performing such procedures with a reduced inflammatory load reduces side effects and can enhance the effectiveness of such types of therapy.

[0036] Each plasmapheresis device can be optimized in various ways, but the device illustrated in Figure 3 includes three different columns 3008a-c. They are identified individually. While one of them will selectively remove gal-3, as discussed above, the other two, in turn, can selectively remove other components, the removal of which can provide therapeutic benefits. Each of these columns 3008 will be provided with its own supporting elements. The associated support elements 3014 may be distributed throughout the length of the tube of the plasmapheresis device. As noted above, the removal and addition of various factors, as well as sampling, will occur in most devices. This is most easily done through discrete support elements 3014, which can be combined with columns 3008 or be separate, as shown in Figure 3.

[0037] Monitoring can be performed by an operator on a control interface 3018, which, as shown, is in effective interaction with the devices and ports described above. The interaction can be hardware, in a manner similar to surgical devices, such as a DaVinci ™ device, or remote and can be achieved using a blue-tooth tool or the Internet. Computer control via the 3018 interface, which traditionally includes a visual monitor that can show a number of essential data, provides the pre-programmed operation of most of the plasmapheresis device of the present invention and the use of alarms and similar methods to alert the operator in situations that arise outside the “normal” parameters . Advantageously, due to the servo control, the operator, either through pre-programmed operations or directly, can act on changing the column, modifying the flow, opening and closing ports, removing and adding, without the need for additional people. This not only limits the cost and processing time, but reduces complexity and limits the risk of infections.

[0038] After a detailed description of many embodiments of the present invention, it should be understood that modifications and variations are possible without going beyond the scope of the legal claims of the invention defined in the attached claims. Moreover, it should be understood that all examples in the present disclosure, although they illustrate many embodiments of the invention, are provided as non-limiting examples and, therefore, should not be construed as limiting the various aspects illustrated in this way.

[0039] Although the present invention is disclosed with reference to certain embodiments, a number of modifications, alterations, and changes to the described embodiments are possible without departing from the scope and scope of the legal claims of the present invention as defined in the attached claims. Accordingly, it is assumed that the present invention is not limited to the described embodiments, but that it has the full scope of legal claims defined by the language of the following claims and their equivalents.

Claims (20)

1. A device for plasmapheresis, containing: the first tube through which the blood removed from the patient of the mammal, moves under the influence of a pump; a separator for separating the cellular components of the specified blood from plasma; a column device through which said plasma flows, said column device selectively removing galectin-3 (gal-3) and other substances associated with galectin-3, including TNF receptor blockers, from said plasma to provide treated plasma; a second tube that receives said treated plasma after passing through said column device and in which said treated plasma is combined with blood cells separated in said separator before returning to said patient; wherein said first and second tubes provide a substantially continuous flow of said blood and plasma from said patient and returning to said mammal a patient, where the specified device contains at least one additional column device for removing a plasma component that is not gal-3. 2. The device according to claim 1, wherein said device contains at least one port through which the therapeutic agent can be added to the specified plasma being treated, separated blood, or both before returning to the specified patient. 3. The device according to claim 1, wherein said device further comprises a monitoring station for providing monitor monitoring and regulation of said plasmapheresis. 4. The device according to claim 3, in which said monitoring station provides for automated control of said separator and said core device and further provides for monitor monitoring of patient parameters selected from the group consisting of pulse rate, oxygen saturation, ECG rhythm, and blood pressure. 5. The device according to claim 1, in which the specified separator is a centrifugal separator. 6. The device according to claim 5, in which the specified centrifuge separator is a centrifuge separator in a continuous stream. 7. The device according to claim 5, in which the specified separator is a separator in a discontinuous flow. 8. The device according to claim 1, wherein said separator is a membrane plasma separator. 9. The device according to claim 1, wherein said device further comprises at least one pump for advancing said cellular components along said tube. 10. The device according to claim 1, additionally containing at least one port through which a blood component selected from the group consisting of B-cells, T-cells, platelets, stem cells and their mixtures can be taken for modification, and then returned to the specified patient on the specified tube. 11. The device according to claim 1, wherein said core device comprises a tube provided with an inner part, wherein the inner part carries a composition that helps an agent that binds gal-3, and wherein said plasma in said tube passes through a portion of said tube which contains the specified agent. 12. The device according to claim 11, in which the specified agent includes a ligand that binds gal-3. 13. The device according to claim 1, wherein said device comprises an additional column device that removes tumor necrosis factor-α, and an additional column device that removes transforming growth factor-β. 14. The device according to claim 9, in which the specified device contains a port before entering the specified separator to add anticoagulant.

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